Songbird: a unique animal model for studying the molecular basis of disorders of vocal development and communication

Like humans, songbirds are one of the few animal groups that learn vocalization. Vocal learning requires coordination of auditory input and vocal output using auditory feedback to guide one’s own vocalizations during a specific developmental stage known as the critical period. Songbirds are good animal models for understand the neural basis of vocal learning, a complex form of imitation, because they have many parallels to humans with regard to the features of vocal behavior and neural circuits dedicated to vocal learning. In this review, we will summarize the behavioral, neural, and genetic traits of birdsong. We will also discuss how studies of birdsong can help us understand how the development of neural circuits for vocal learning and production is driven by sensory input (auditory information) and motor output (vocalization).     

Deafening drives cell-type-specific changes to dendritic spines in a sensorimotor nucleus important to learned vocalizations

Hearing loss prevents vocal learning and causes learned vocalizations to deteriorate, but how vocalization-related auditory feedback acts on neural circuits that control vocalization remains poorly understood. We deafened adult zebra finches, which rely on auditory feedback to maintain their learned songs, to test the hypothesis that deafening modifies synapses on neurons in a sensorimotor nucleus important to song production. Longitudinal in vivo imaging revealed that deafening selectively decreased the size and stability of dendritic spines on neurons that provide input to a striatothalamic pathway important to audition-dependent vocal plasticity, and changes in spine size preceded and predicted subsequent vocal degradation. Moreover, electrophysiological recordings from these neurons showed that structural changes were accompanied by functional weakening of both excitatory and inhibitory synapses, increased intrinsic excitability, and changes in spontaneous action potential output. These findings shed light on where and how auditory feedback acts within sensorimotor circuits to shape learned vocalizations.     

A bird's eye view: top down intracellular analyses of auditory selectivity for learned vocalizations

The "song system" refers to a group of interconnected brain nuclei necessary for the utterance of learned song and for the generation of vocal plasticity important to both song learning and adult song maintenance. Although song learning and, in some species, song maintenance depend on auditory feedback, how audition influences vocalization remains unknown. One attractive idea is that auditory signals propagate directly to those telencephalic nuclei implicated in song patterning, providing a convenient substrate for sensorimotor integration. Consistent with this idea, auditory neurons highly selective for the bird's own song have been detected in telencephalic song nuclei, and lesions of these structures can impair song perception as well as song production. This review discusses evidence for an auditory-perceptual role of the song system, the anatomical pathways by which auditory information enters the song system, the synaptic events underlying highly selective action potential responses to learned song, and the possible roles such activity could play in song learning and maintenance.     

A Lightweight,Headphones-based System for Manipulating Auditory Feedback in Songbirds

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and sensorimotor plasticity¹. Consistent sensory perturbations result in compensatory changes in motor output, reflecting changes in feedforward motor control that reduce the experienced feedback error. By quantifying how different sensory feedback errors affect human behavior, prior studies have explored how visual signals are used to recalibrate arm movements^2,3 and auditory feedback is used to modify speech production^4-7. The strength of this approach rests on the ability to mimic naturalistic errors in behavior, allowing the experimenter to observe how experienced errors in production are used to recalibrate motor output.Songbirds provide an excellent animal model for investigating the neural basis of sensorimotor control and plasticity^8,9. The songbird brain provides a well-defined circuit in which the areas necessary for song learning are spatially separated from those required for song production, and neural recording and lesion studies have made significant advances in understanding how different brain areas contribute to vocal behavior^9-12. However, the lack of a naturalistic error-correction paradigm - in which a known acoustic parameter is perturbed by the experimenter and then corrected by the songbird - has made it difficult to understand the computations underlying vocal learning or how different elements of the neural circuit contribute to the correction of vocal errors¹³.The technique described here gives the experimenter precise control over auditory feedback errors in singing birds, allowing the introduction of arbitrary sensory errors that can be used to drive vocal learning. Online sound-processing equipment is used to introduce a known perturbation to the acoustics of song, and a miniaturized headphones apparatus is used to replace a songbird''s natural auditory feedback with the perturbed signal in real time. We have used this paradigm to perturb the fundamental frequency (pitch) of auditory feedback in adult songbirds, providing the first demonstration that adult birds maintain vocal performance using error correction¹⁴. The present protocol can be used to implement a wide range of sensory feedback perturbations (including but not limited to pitch shifts) to investigate the computational and neurophysiological basis of vocal learning.     

Birdsong: From behaviour to brain

Although vocal communication is wide-spread in animal kingdom, the use of learned (in contrast to innate) vocalization is very rare. We can find it only in few animal taxa: human, bats, whales and dolphins, elephants, parrots, hummingbirds, and songbirds. There are several parallels between human and songbird perception and production of vocal signals. Hence, many studies take interest in songbird singing for investigating the neural bases of learning and memory. Brain circuits controlling song learning and maintenance consist of two pathways — a vocal motor pathway responsible for production of learned vocalizations and anterior forebrain pathway responsible for learning and modifying the vocalizations. This review provides an overview of the song organization, its behavioural traits, and neural regulations. The recently expanding area of molecular mapping of the behaviour-driven gene expression in brain represents one of the modern approaches to the study the function of vocal and auditory areas for song learning and maintenance in birds.     

Development of Auditory-Vocal Perceptual Skills in Songbirds

Songbirds are one of the few groups of animals that learn the sounds used for vocal communication during development. Like humans, songbirds memorize vocal sounds based on auditory experience with vocalizations of adult “tutors”, and then use auditory feedback of self-produced vocalizations to gradually match their motor output to the memory of tutor sounds. In humans, investigations of early vocal learning have focused mainly on perceptual skills of infants, whereas studies of songbirds have focused on measures of vocal production. In order to fully exploit songbirds as a model for human speech, understand the neural basis of learned vocal behavior, and investigate links between vocal perception and production, studies of songbirds must examine both behavioral measures of perception and neural measures of discrimination during development. Here we used behavioral and electrophysiological assays of the ability of songbirds to distinguish vocal calls of varying frequencies at different stages of vocal learning. The results show that neural tuning in auditory cortex mirrors behavioral improvements in the ability to make perceptual distinctions of vocal calls as birds are engaged in vocal learning. Thus, separate measures of neural discrimination and behavioral perception yielded highly similar trends during the course of vocal development. The timing of this improvement in the ability to distinguish vocal sounds correlates with our previous work showing substantial refinement of axonal connectivity in cortico-basal ganglia pathways necessary for vocal learning.     

Is the Capacity for Vocal Learning in Vertebrates Rooted in Fish Schooling Behavior?

The capacity to learn and reproduce vocal sounds has evolved in phylogenetically distant tetrapod lineages. Vocal learners in all these lineages express similar neural circuitry and genetic factors when perceiving, processing, and reproducing vocalization, suggesting that brain pathways for vocal learning evolved within strong constraints from a common ancestor, potentially fish. We hypothesize that the auditory-motor circuits and genes involved in entrainment have their origins in fish schooling behavior and respiratory-motor coupling. In this acoustic advantages hypothesis, aural costs and benefits played a key role in shaping a wide variety of traits, which could readily be exapted for entrainment and vocal learning, including social grouping, group movement, and respiratory-motor coupling. Specifically, incidental sounds of locomotion and respiration (ISLR) may have reinforced synchronization by communicating important spatial and temporal information between school-members and extending windows of silence to improve situational awareness. This process would be mutually reinforcing. Neurons in the telencephalon, which were initially involved in linking ISLR with forelimbs, could have switched functions to serve vocal machinery (e.g. mouth, beak, tongue, larynx, syrinx). While previous vocal learning hypotheses invoke transmission of neurons from visual tasks (gestures) to the auditory channel, this hypothesis involves the auditory channel from the onset. Acoustic benefits of locomotor-respiratory coordination in fish may have selected for genetic factors and brain circuitry capable of synchronizing respiratory and limb movements, predisposing tetrapod lines to synchronized movement, vocalization, and vocal learning. We discuss how the capacity to entrain is manifest in fish, amphibians, birds, and mammals, and propose predictions to test our acoustic advantages hypothesis.     

At the interface of the auditory and vocal motor systems: NIf and its role in vocal processing,production and learning

Communication between auditory and vocal motor nuclei is essential for vocal learning. In songbirds, the nucleus interfacialis of the nidopallium (NIf) is part of a sensorimotor loop, along with auditory nucleus avalanche (Av) and song system nucleus HVC, that links the auditory and song systems. Most of the auditory information comes through this sensorimotor loop, with the projection from NIf to HVC representing the largest single source of auditory information to the song system. In addition to providing the majority of HVC’s auditory input, NIf is also the primary driver of spontaneous activity and premotor-like bursting during sleep in HVC. Like HVC and RA, two nuclei critical for song learning and production, NIf exhibits behavioral-state dependent auditory responses and strong motor bursts that precede song output. NIf also exhibits extended periods of fast gamma oscillations following vocal production. Based on the converging evidence from studies of physiology and functional connectivity it would be reasonable to expect NIf to play an important role in the learning, maintenance, and production of song. Surprisingly, however, lesions of NIf in adult zebra finches have no effect on song production or maintenance. Only the plastic song produced by juvenile zebra finches during the sensorimotor phase of song learning is affected by NIf lesions. In this review, we carefully examine what is known about NIf at the anatomical, physiological, and behavioral levels. We reexamine conclusions drawn from previous studies in the light of our current understanding of the song system, and establish what can be said with certainty about NIf’s involvement in song learning, maintenance, and production. Finally, we review recent theories of song learning integrating possible roles for NIf within these frameworks and suggest possible parallels between NIf and sensorimotor areas that form part of the neural circuitry for speech processing in humans.     

听觉皮层信号处理

听觉系统和视觉系统的不同之处在于：听觉系统在外周感受器和听皮层间具有更长的皮层下通路和更多的突触联系。该特殊结构反应了听觉系统从复杂听觉环境中提取与行为相关信号的机制与其他感觉系统不同。听皮层神经信号处理包括两种重要的转换机制,声音信号的非同构转换以及从声音感受到知觉层面的转换。听觉皮层神经编码机制同时也受到听觉反馈和语言或发声过程中发声信号的调控。听觉神经科学家和生物医学工程师所面临的挑战便是如何去理解大脑中这些转换的编码机制。我将会用我实验室最近的一些发现来阐述听觉信号是如何在原听皮层中进行处理的,并讨论其对于言语和音乐在大脑中的处理机制以及设计神经替代装置诸如电子耳蜗的意义。我们使用了结合神经电生理技术和量化工程学的方法来研究这些问题。     

Auditory responses in the vocal motor system of budgerigars

Budgerigars (Melopsittacus undulatus) are small Australian parrots that can imitate novel sounds in adulthood and therefore serve as a convenient model system for the study of vocal learning in adult animals. Previous anatomical studies had indicated that known auditory regions in the telencephalon of budgerigars are connected, albeit indirectly and rather sparsely, to vocal motor nuclei. Physiological evidence for connections between the auditory and vocal motor systems in budgerigars had been lacking, however. Here, we show that neurons in a telencephalic vocal motor region, i.e., the central nucleus of the lateral neostriatum (NLc), are responsive to auditory stimuli in isoflurane-anesthetized budgerigars. These responses are highly variable from trial to trial and frequently have latencies in excess of 100 ms. Neurons in NLc generally respond better to a budgerigar's own contact call than to a white noise stimulus, but the response preferences of NLc neurons in budgerigars are generally weaker and more diverse than the response preferences of neurons in the high vocal center of songbirds, which is probably analogous to NLc. These data indicate that parrots and songbirds, which have evolved the ability to learn vocalizations independently of one another, have both evolved physiologically effective connections between their auditory and vocal motor systems. Interestingly, however, the anatomical pathways by which the auditory and vocal motor systems interact, and the physiological details of how they communicate, appear to be significantly different between the two taxa.     

LMAN lesions prevent song degradation after deafening without reducing HVC neuron addition

In some songbirds perturbing auditory feedback can promote changes in song structure well beyond the end of song learning. One factor that may drive vocal change in such deafened birds is the ongoing addition of new vocal-motor neurons into the song system. Without auditory feedback to guide their incorporation, the addition of these new neurons could disrupt the established song pattern. To assess this hypothesis, the authors determined if neuronal recruitment into the vocal motor nucleus HVC is affected by neural signals that influence vocal change in adult deafened birds. Such signals appear to be conveyed via LMAN, a nucleus in the anterior forebrain that is necessary for vocal change after deafening. Here the authors tested whether LMAN lesions might restrict song degradation after deafening by reducing the addition or survival of new HVC neurons that would otherwise corrupt the ongoing song pattern. Using [3H]thymidine autoradiography to identify neurons generated in adult zebra finches, it was shown here that LMAN lesions do not reduce the number or percent of new HVC neurons surviving for either several weeks or months after [3H]thymidine labeling. However, the authors confirmed previous reports that LMAN lesions restrict vocal change after deafening. These data suggest that neurons incorporated into the adult HVC may form behaviorally adaptive connections without requiring auditory feedback, and that any role such neurons may play in promoting vocal change after adult deafening requires anterior forebrain pathway output.     

Auditory feedback in learning and maintenance of vocal behaviour

Songbirds are one of the best-studied examples of vocal learners. Learning of both human speech and birdsong depends on hearing. Once learned, adult song in many species remains unchanging, suggesting a reduced influence of sensory experience. Recent studies have revealed, however, that adult song is not always stable, extending our understanding of the mechanisms involved in song maintenance, and their similarity to those active during song learning. Here we review some of the processes that contribute to song learning and production, with an emphasis on the role of auditory feedback. We then consider some of the possible neural substrates involved in these processes, particularly basal ganglia circuitry. Although a thorough treatment of human speech is beyond the scope of this article, we point out similarities between speech and song learning, and ways in which studies of these disparate behaviours complement each other in developing an understanding of general principles that contribute to learning and maintenance of vocal behaviour.     

Temporal recalibration in vocalization induced by adaptation of delayed auditory feedback

Background

We ordinarily perceive our voice sound as occurring simultaneously with vocal production, but the sense of simultaneity in vocalization can be easily interrupted by delayed auditory feedback (DAF). DAF causes normal people to have difficulty speaking fluently but helps people with stuttering to improve speech fluency. However, the underlying temporal mechanism for integrating the motor production of voice and the auditory perception of vocal sound remains unclear. In this study, we investigated the temporal tuning mechanism integrating vocal sensory and voice sounds under DAF with an adaptation technique.

Methods and Findings

Participants produced a single voice sound repeatedly with specific delay times of DAF (0, 66, 133 ms) during three minutes to induce ‘Lag Adaptation’. They then judged the simultaneity between motor sensation and vocal sound given feedback. We found that lag adaptation induced a shift in simultaneity responses toward the adapted auditory delays. This indicates that the temporal tuning mechanism in vocalization can be temporally recalibrated after prolonged exposure to delayed vocal sounds. Furthermore, we found that the temporal recalibration in vocalization can be affected by averaging delay times in the adaptation phase.

Conclusions

These findings suggest vocalization is finely tuned by the temporal recalibration mechanism, which acutely monitors the integration of temporal delays between motor sensation and vocal sound.     

Sensitive periods and circuits for learned birdsong

Experience influences the development of certain behaviors and their associated neural circuits during a discrete period after birth. Songbirds, with their highly quantifiable vocal output and well-delineated vocal control circuitry, provide an excellent context in which to examine the neural mechanisms regulating sensitive periods for learning. Recent discoveries indicate that auditory input to the vocal control circuitry in songbirds is dynamically modulated and show that neural circuitry previously thought to be used only in plastic juvenile song may also actively maintain stable adult song. These findings provide important clues to how sensitive periods for auditory feedback and vocal plasticity are regulated during song development.     

Of Mice,Birds, and Men: The Mouse Ultrasonic Song System Has Some Features Similar to Humans and Song-Learning Birds

Humans and song-learning birds communicate acoustically using learned vocalizations. The characteristic features of this social communication behavior include vocal control by forebrain motor areas, a direct cortical projection to brainstem vocal motor neurons, and dependence on auditory feedback to develop and maintain learned vocalizations. These features have so far not been found in closely related primate and avian species that do not learn vocalizations. Male mice produce courtship ultrasonic vocalizations with acoustic features similar to songs of song-learning birds. However, it is assumed that mice lack a forebrain system for vocal modification and that their ultrasonic vocalizations are innate. Here we investigated the mouse song system and discovered that it includes a motor cortex region active during singing, that projects directly to brainstem vocal motor neurons and is necessary for keeping song more stereotyped and on pitch. We also discovered that male mice depend on auditory feedback to maintain some ultrasonic song features, and that sub-strains with differences in their songs can match each other''s pitch when cross-housed under competitive social conditions. We conclude that male mice have some limited vocal modification abilities with at least some neuroanatomical features thought to be unique to humans and song-learning birds. To explain our findings, we propose a continuum hypothesis of vocal learning.     

Auditory–vocal mirroring in songbirds

Mirror neurons are theorized to serve as a neural substrate for spoken language in humans, but the existence and functions of auditory–vocal mirror neurons in the human brain remain largely matters of speculation. Songbirds resemble humans in their capacity for vocal learning and depend on their learned songs to facilitate courtship and individual recognition. Recent neurophysiological studies have detected putative auditory–vocal mirror neurons in a sensorimotor region of the songbird''s brain that plays an important role in expressive and receptive aspects of vocal communication. This review discusses the auditory and motor-related properties of these cells, considers their potential role on song learning and communication in relation to classical studies of birdsong, and points to the circuit and developmental mechanisms that may give rise to auditory–vocal mirroring in the songbird''s brain.     

Sensory-motor interactions modulate a primate vocal behavior: antiphonal calling in common marmosets

A fundamental issue in neuroscience pertains to how different cortical systems interact to generate behavior. One of the most direct ways to address this issue is to investigate how sensory information is encoded and used to produce a motor response. Antiphonal calling is a natural vocal behavior that involves individuals producing their species-specific long distance vocalization in response to hearing the same call and engages both the auditory and motor systems, as well as the cognitive neural systems involved in decision making and categorization. Here we present results from a series of behavioral experiments investigating the auditory–vocal interactions during antiphonal calling in the common marmoset (Callithrix jacchus). We manipulated sensory input by placing subjects in different social contexts and found that the auditory input had a significant effect on call timing and propensity to call. Playback experiments tested the significance of the timing of vocal production in antiphonal calling and showed that a short latency between antiphonal calls was necessary to maintain reciprocal vocal interactions. Overall, this study shows that sensory-motor interactions can be experimentally induced and manipulated in a natural primate vocal behavior. Antiphonal calling represents a promising model system to examine these issues in non-human primates at both the behavioral and neural levels.     

Children's development of self-regulation in speech production

Species-specific vocalizations fall into two broad categories: those that emerge during maturation, independent of experience, and those that depend on early life interactions with conspecifics. Human language and the communication systems of a small number of other species, including songbirds, fall into this latter class of vocal learning. Self-monitoring has been assumed to play an important role in the vocal learning of speech and studies demonstrate that perception of your own voice is crucial for both the development and lifelong maintenance of vocalizations in humans and songbirds. Experimental modifications of auditory feedback can also change vocalizations in both humans and songbirds. However, with the exception of large manipulations of timing, no study to date has ever directly examined the use of auditory feedback in speech production under the age of 4. Here we use a real-time formant perturbation task to compare the response of toddlers, children, and adults to altered feedback. Children and adults reacted to this manipulation by changing their vowels in a direction opposite to the perturbation. Surprisingly, toddlers' speech didn't change in response to altered feedback, suggesting that long-held assumptions regarding the role of self-perception in articulatory development need to be reconsidered.     

黄喉鵐高级发声中枢及其壳区的神经投射和P物质在发声听觉中枢的分布

用生物素示踪法和P物质 (SP)免疫组化技术研究表明 :黄喉的高级发声中枢 (HVc)接受端脑听区 (L)、新纹状体中部界面核、新纹状体巨细胞核 (MAN)、丘脑葡萄形核、桥脑蓝斑核的传入 ,并有神经纤维投射到古纹状体栎核 (RA)和嗅叶X区 (X) ;HVc壳投射到RA壳并接受L的传入。听觉控制与学习通路与发声中枢之间有许多神经联系 ,提示黄喉发声学习依赖于听觉反馈。在HVc、RA和MAN有SP阳性细胞体 ,在X、中脑背内侧核和延髓舌下神经核气管鸣管部、丘脑卵圆核壳区、中脑背外侧核壳区及中脑丘间核有SP阳性纤维和终末。SP广泛分布于发声 -听觉中枢 ,可能参与了它们的活动     

Neural auditory selectivity develops in parallel with song

The zebra finch learns his song by memorizing a tutor's vocalization and then using auditory feedback to match his current vocalization to this memory, or template. The neural song system of adult and young birds responds to auditory stimuli, and exhibits selective tuning to the bird's own song (BOS). We have directly examined the development of neural tuning in the song motor system. We measured song system responses to vocalizations produced at various ages during sleep. We now report that the auditory response of the song motor system and motor output are linked early in song development. During sleep, playback of the current BOS induced a response in the song nucleus HVC during the song practice period, even when the song consisted of little more than repeated begging calls. Halfway through the sensorimotor period when the song was not yet in its final form, the response to BOS already exceeded that to all other auditory stimuli tested. Moreover, responses to previous, plastic versions of BOS decayed over time. This indicates that selective tuning to BOS mirrors the vocalization that the bird is currently producing.